Artesunate Enhances the Cytotoxicity of 5-Aminolevulinic Acid-Based Sonodynamic Therapy against Mouse Mammary Tumor Cells In Vitro.

Sonodynamic therapy (SDT) kills tumor cells through the synergistic effects of ultrasound (US) and a sonosensitizer agent. 5-Aminolevulinic acid (5-ALA) has been used as a sonodynamic sensitizer for cancer treatment. However, studies have shown that 5-ALA-based SDT has limited efficacy against malignant tumors. In this study, we examined whether artesunate (ART) could enhance the cytotoxicity of 5-ALA-based SDT against mouse mammary tumor (EMT-6) cells in vitro. In the ART, ART + US, ART + 5-ALA, and ART + 5-ALA + US groups, the cell survival rate correlated with ART concentration, and decreased with increasing concentrations of ART. Morphologically, many apoptotic and necrotic cells were observed in the ART + 5-ALA + US group. The percentage of reactive oxygen species-positive cells in the ART + 5-ALA + US group was also significantly higher than that in the 5-ALA group (p = 0.0228), and the cell death induced by ART + 5-ALA + US could be inhibited by the antioxidant N-acetylcysteine. These results show that ART offers great potential in enhancing the efficacy of 5-ALA-based SDT for the treatment of cancer. However, these results are only based on in vitro studies, and further in vivo studies are required.

Factors Associated with Number of Present Teeth in Adults in Japanese Urban City.

The purpose of this study was to identify which factors were associated with number of present teeth (PT) in older adults as their clarification might contribute to maintaining the number of PT in this population. These factors were investigated by means of a questionnaire on oral health condition, general health behavior, and anamnesis of diabetes mellitus together with a dental examination. Data were obtained from a total of 7,741 Japanese subjects (2,623 males and 5,118 females), all of whom were aged 40, 50, or 60 years. Significant differences were observed in the mean number of PT between a positive or negative response to questions on anamnesis of diabetes mellitus, current smoking, and having loose teeth in all age groups in males; significant differences were also observed for current smoking, lower intake of dairy milk products, having loose teeth, no experience of tooth brushing instruction, and poor oral hygiene status in all age groups in females. Stepwise logistic regression analysis revealed having loose teeth (odds ratio [OR], 1.82), gingival swelling (OR, 1.40), an anamnesis of diabetes mellitus (OR, 1.72), current smoking (OR, 1.86), lower intake of dairy milk products (OR, 1.22), preference for salty seasonings (OR, 1.23), frequent sweet intake (OR, 1.29), no experience of tooth brushing instruction (OR, 1.38), and poor oral hygiene (OR, 2.04) as significant risk factors for the number of PT being <24 after adjusting for age and sex. These results indicate that an anamnesis of diabetes mellitus, a history of smoking, the presence of loose teeth, and poor oral hygiene status in a self-reported evaluation of oral health might correlate with <24 PT in older adults.

Relationship between General Health, Lifestyle, Oral Health, and Periodontal Disease in Adults: A Large Cross-sectional Study in Japan.

The aim of this study was to investigate how general health, oral conditions, and lifestyle were associated with periodontal disease in adults, as clarifying this relationship may be useful in preventing periodontal disease. Medical checkups were conducted on individuals aged 40, 50, or 60 years. Data were obtained for analysis on a total of 36,110 patients (men, 12,784; women, 22,896). A stepwise logistic regression model was used to calculate the odds ratio (OR) for patients who were ≥code 3 according to the Community Periodontal Index (CPI). Approximately 40, 60, and 70% of men aged 40, 50, and 60 years, respectively, had a CPI score of ≥3. There were 10% fewer women than men at each age. Stepwise logistic regression revealed a BMI score of ≥30 kg/m2(OR, 1.44; 95% confidence interval [95%CI], 1.20-1.73); systolic blood pressure of ≥140 mmHg (OR, 1.09; 95%CI, 1.02-1.18); a fasting blood sugar level of ≥110 mg/dl (OR, 1.17; 95%CI, 1.04-1.30); high-density lipoprotein cholesterol level of <40 mg/dl (OR, 1.21; 95%CI, 1.06-1.37); smoker (OR, 1.59; 95%CI, 1.48-1.71); drinking ≥3 cups of Japanese sake per day (OR, 1.09; 95%CI, 1.05-1.14); use of salts for seasoning (OR, 1.17; 95%CI, 1.07-1.28); and fair and poor oral hygiene (OR, 2.27; 95%CI, 2.08-2.47) as significant risk factors for a CPI score of ≥3. These results suggest that smoking, oral hygiene status, and factors associated with metabolic syndrome are associated with periodontitis. This indicates that health guidance on tooth brushing, the importance of quitting smoking, and control of obesity may be effective in preventing the development of periodontal disease in adults.

Bleomycin enhances the efficacy of sonodynamic therapy using aluminum phthalocyanine disulfonate.

Sonodynamic therapy (SDT), or ultrasound combined with sonosensitization, is a promising approach because it is noninvasive and penetrates deeper than light does in photodynamic therapy. We examined whether bleomycin (BLM) could improve the efficacy of SDT. We performed an in vitro study using Colon-26 cells, which are derived from mouse colon cancer. SDT with BLM was significantly more cytotoxic than SDT alone both in vitro and in vivo. We also observed an ultrasound intensity-dependent cytotoxic effect of SDT with BLM. These findings suggest that SDT with BLM might provide a novel noninvasive treatment for deep-seated tumors.

Sonodynamic therapy using 5-aminolevulinic acid enhances the efficacy of bleomycin.

Sonodynamic therapy (SDT) kills tumor cells through the synergistic effects of ultrasound and a sonosensitizer agent. We examined whether 5-aminolevulinic acid (5-ALA)-based SDT at 1 or 3 MHz could enhance the cytotoxicity of bleomycin (BLM) toward mouse mammary tumor cells both in vitro and in vivo. At 1 MHz, cell viability in the 5-ALA-based SDT group at 1, 2, and 3 W/cm(2) was 34.30%, 50.90%, and 60.16%, respectively. Cell viability in the 5-ALA-based SDT+BLM group at 1, 2, and 3 W/cm(2) was 0.09%, 0.32%, and 0.17%, respectively. In contrast, at 3 MHz, 5-ALA-based SDT+BLM did not show pronounced cytotoxicity. In the in vivo study, 5-ALA-based SDT+BLM was significantly more cytotoxic than 5-ALA-based SDT at 1 MHz and 3 MHz. These findings suggest that the mechanism of tumor shrinkage induced by 5-ALA-based SDT+BLM might involve not only direct cell killing, but also vascular shutdown. Thus, we show here that 5-ALA-based SDT enhances the efficacy of BLM both in vitro and in vivo.

5-Aminolevulinic Acid Enhances Ultrasound-mediated Antitumor Activity via Mitochondrial Oxidative Damage in Breast Cancer.

BACKGROUND/AIM: 5-Aminolevulinic acid (5-ALA), a precursor of protoporphyrin IX (PpIX), is now used for photodynamic therapy (PDT) of pre-cancers of the skin and photodynamic diagnosis (PDD) of brain tumors. Sonodynamic therapy (SDT) of cancers with ultrasound has been studied using 5-ALA as a sonosensitizer. In this article, we evaluated the sonosensitizing activity and mode of action of 5-ALA/PpIX by using mouse mammary tumor EMT6 cells. RESULTS: 5-ALA-SDT showed significant antitumor effects toward EMT6 cells in vitro and in vivo. The fluorescence of MitoSOX Red, an indicator specific for mitochondrial superoxide, was significantly increased by 5-ALA-SDT. Moreover, the fluorescence derived from JC-1, an indicator of mitochondrial membrane potential, was also significantly increased by 5-ALA-SDT. These findings suggest that mitochondria are one of the target organelles of 5-ALA-SDT. PpIX enhanced reactive oxygen species (ROS) production from tert-butyl hydroperoxide (tBHP), suggesting that PpIX might stabilize or promote ROS generation from tBHP. CONCLUSION: 5-ALA-SDT showed an antitumor effect in mouse mammary tumor EMT6 cells through oxidation of the mitochondrial membrane via ROS production.